...composed of four peptide chains (two identical heavy chains and two identical light chains) each with a constant...antibodies means that it is relatively easy to physically divide up the different regions, both through... as secondary antibodies, have much broader applicability. It is fairly easy to find these in a Fab fragment...fragments described above can be used for similar applications and have similar advantages, though with some...David vs. Goliath: The Structure, Function, and Clinical Prospects of Antibody Fragments. Antibodies 8(...

...problems. Rapidly dividing cells initiate genome replication more than once per cell division and thus, genes...genes closer to the origin of replication will be overrepresented compared to those further away. As a...cells are dividing rapidly, multiple rounds of replication are initiated before the cell has divided. This...maximal gene expression near genomic origins of replication, the stabilized promoters created by Segall-Shapiro...the control of IPTG. Because RepA is a plasmid replication initiator protein, they could alter copy number...dosage in E. coli using DIAL strains." Journal of biological engineering 5.1 (2011): 10. PubMed PMID: 21787416...

...editing performance at therapeutically relevant gene targets and expand the application to the possible treatment...AsCas12f1. They found that this system, without any modifications, is effective in genome editing in both bacteria...ribonucleoprotein, and AAV.  Read the paper in Nature Chemical Biology Find the AsCas12f1 plasmids   Selective... Mike Lacy The retroviral-like protein PEG10 specifically binds its own mRNA to facilitate its packaging... the prime editor, yielding PEmax, to work synergistically with epegRNAs and PE4 and PE5. These enhanced...

... audience and convey your ideas clearly and strategically is powerful in almost any career. It allows ...interview better! As a scientist turned science communicator, I’ve done a lot of public speaking, and I'm... the open windows in the middle of your talk. Technical difficulties turn your slide deck into a chalk...everyone! Everybody makes mistakes, encounters technical difficulties, and runs into unexpected stumbling..., to a group of peers who are there to provide critical feedback, often with a semi-international audience...your own native language. And, at least in the biomedical field, with very little text on the slides and...

...days prior to the implementation date, our data indicated that, while 69% of all postdocs were expected ... department at the University of Massachusetts Medical School. We are working with postdocs at other institutions...sent, with the lack of transparency and poor communication of their plans (many postdocs - and in some ...to reach out to institutions with a statement indicating that we encourage them to keep raising salaries... December 1st. Salary inconsistencies across geographical regions When we originally started gathering...changes in science. Follow him on Twitter @BiophysicalFrog and Future of Research @FORsymp.   Additional...

...and on self-propagation by reproduction. Both physiological processes are controlled by deliberate and compulsory...receptors (NRs), their coregulators, and their physiological endocrine ligands. Nuclear receptors and their...murine members. NRs permit the integration and communication of such signals between central and peripheral...circadian regulation, as well as in important physiological states such as adipogenesis and macrophage activation... activation. Our mass spectrometry-affinity purification proteomic profiling of coregulators in HeLa cells...widely used today.  The sheer abundance of these critical research tools is a double-edged sword, however...

... for PAM-less editing   Guides for CRISPR are typically selected by the ‘quality’ of the gRNA sequence...location of the cut and subsequent edit. In these applications, location takes priority over cutting efficiency...the Liu lab through phage-assisted evolution, specifically with the goal of generating Cas enzymes with...compatibility. This alternative PAM requirement adds a significant amount of flexibility compared to a strict single-compatibility...in double stranded cleavage and base editing applications, with high editing capacity in both contexts...increases the flexibility of gRNA placement significantly over SpCas9 and more so than Cas9s with an NG...

...prevent the formation of replication competent lentivirus (RCL). Replication competence can arise from...end-users, size of the transgene, and downstream applications should all be considered when choosing the correct...unconcentrated prep. For example, at Addgene we typically concentrate 100-fold meaning that a 10 cm2 dish... as resources available, cost and downstream applications. For example, while the PEG precipitation method...lentivirus is the cell line that is used for titering. Typically researchers will choose a relatively permissive...

.... When an ELISA kit is ordered, the wells are typically already coated with the antibody. Then, you add...process is the addition of a detection antibody (typically a monoclonal antibody) that also recognizes the...the most common type of competitive ELISA is a modification of the sandwich ELISA as described here, the...ELISA, the only difference being the additional amplification step, which helps to visualize low-abundance...concentrations of your protein of interest. The optical density (OD) - the measure of light absorbed by...concentration of any experimental wells in that range of optical densities. If your standard curve does not have...

...targeting anywhere in an exon. But things get more complicated if you’d like to use homology directed repair...edits or knock-ins. Editing efficiency decreases drastically once your cut site is more than 10 bp away from...They created a selection phage (SP) to carry catalytically dead SpCas9 (dCas9) fused to the bacterial polymerase...loci displayed a broad range of PAM sequences, indicating they may be less predictable than SpCas9 off-... that we have only scratched the surface of modifications that can be made to Cas9. It is a pleasant surprise...editing rates. This PAM flexibility should expand applications that require very specific targeting, like homology...

...Gonçalves and his colleagues at Leiden University Medical Center are now available at Addgene along with ...and very efficient introduction of DNA into therapeutically relevant, non-transformed mammalian cells. ...and quiescent, post-mitotic mammalian cells. Applications of CRISPR delivery by adenoviral vectors Since...and used AdV vectors to edit genes in several biological systems. It has been used for instance in inherited...Strategies for Tissue-Specific Targeting. In: Applications of viruses for cancer therapy. Elsevier, pp ...Viral vectors for gene therapy: Current state and clinical perspectives. Biochemistry Moscow 81:700–708 ....

...experts in science communication in real time! I follow great online science communicators like Phil Plait...nontraditional” career path long ago. Use your analytical mind to test and discover what career suits you...about any topic I like and gain experience in communication, editing, marketing, and website organization.... She is particularly interested in science communication and fundamental molecular biology.      Additional...  science communication...

... familial aspect of Addgene. As our size has drastically increased, it would have been a tight squeeze...event! Addgene company culture At Addgene we organically developed the philosophy of  “work hard, play...department) right in your breakroom. 5. Throw some physical activity into the mix It’s easy to sit around ...be an obvious one, but don’t make things too complicated for yourself (especially if you are the sole ...sole event planner)! If you have a very intricate itinerary, there is a higher likelihood that something ...Delegate- it will save you time and sanity!    10. Communicate Make sure people know that there is something...

...Societies, such as the American Society for Cell Biology (ASCB) or the American Chemical Society (ACS) are ...often scarce and difficult to identify. So what specifically should you do? What follows are just a few suggestions... in fact, some of the very things that were so critical to me landing my own position! Conferences One...know all in one place! This information can be critical to a focused and effective job search. As with...

...development happening at Addgene under the hood. A significant portion of our development may not be visible...ourselves. We are also able to consider different technical approaches that may come up during discussion.... information. It is quite possible that the clarifications lead to the story being scrapped or broken ...While writing the code, we may need additional clarification and again we collaborate with the Product Team... been updated in several years) or they make significant changes that make it incompatible with our software...software from Python 2.7 to Python 3.5. There are significant changes to the language and it took us a total...

...regions and allocate specific functions to them. Electrical recordings and lesioning studies have been essential...within the LC of mice. When those neurons are chemogenetically activated, norepinephrine release causes a...test has been used since the 1970s to test the efficacy of antidepressant treatment in rodents. In this...responsible for the antidepressant effect. To specifically activate the Drd1 neurons, they injected AAV2... "Hypothalamic CNTF volume transmission shapes cortical noradrenergic excitability upon acute stress."...long-lasting antidepressant effects." Nature communications 10.1 (2019): 223. PubMed PMID: 30644390. PubMed...

...possess cell walls. As a result, Mycoplasma are intrinsically resistant to this class of antibiotic. Acquired...Microbes (bacteria included) are very skilled at the delicate art of survival. Existing virtually ubiquitously...of intentions. The take-home? Always take your medication as prescribed. How do we know which bacteria ...When antibiotic resistance testing is requested clinically, this involves growing a standardized amount ...-trusted method that is frequently used in the clinical laboratory. What can we do to stop the emergence... is passionate about health equity, science communication, and public health.     References 1. O'Neill...

...FRET during a journal club, my guitarist brain automatically thought about the raised element found on the...allows the detection of molecules' interactions, modifications or dissociations in situ. Used since the mid...interactions in living cells using a microscope. Historically, molecular interactions were detected by indirect...Periasamy defined FRET as "a distance-dependent physical process by which energy is transferred non-radiatively...highest absorbing acceptor and fluorophores with significant overlap in their spectra. The pair CFP-YFP was...14. Zimmermann T,  Terjung S. FRET Basics and Applications: An EAMNET Teaching Module http://www.embl.de...

...resistance gene and the origin of replication are necessary to replicate plasmids and make sure the cells... promoters only work in prokaryotic cells and typically only in the same or closely related species from... be ways to turn them on or off at will with a chemical, heat, or light. In the cell, promoters themselves...scientists have created synthetic promoters, which typically include some combination of other promoter elements...Inducible with Tetracyline or its derivatives.  Typically contains a minimal promoter with low basal activity...

... the planning section below, it’s important to dedicate specific amounts of time to specific tasks beforehand...activities that are needed to accomplish these goals. Practically speaking, this means I write everything in a ...down? It’s more viscerally appealing for me to physically cross things out (it just feels good) and I remember...instance, if I don’t have draft blog post ready for publication for tomorrow, I’d better upload that post from...handle doing a particularly annoying task and then dedicating that much time to it each day. Once you get over...Student Community Learn about Careers in Science Communication Resources at Addgene.org Check out Careers...